The antioxidant drink “effective microorganism‐X (EM‐X)” pre‐treatment attenuates the loss of nigrostriatal dopaminergic neurons in 6‐hydroxydopamine‐lesion rat model of Parkinson's disease

There is continued interest in the assessment and potential use of antioxidants as neuroprotective agents in diseases associated with increased oxidative stress, such as Parkinson's disease. The neuroprotective effect of a natural antioxidant drink, EM‐X (a ferment derivative of unpolished rice, papaya and seaweeds with effective microorganisms), was investigated using the 6‐hydroxydopamine (6‐OHDA)‐lesion rat model of Parkinson's disease. The nigrostriatal dopaminergic neurons were unilaterally lesioned with 6‐OHDA (8 μg) in rats that were treated with a 10‐times diluted EM‐X drink (dilEM‐X), standard EM‐X drink (stdEM‐X) or tap water for 4 days. Seven days post lesion, the integrity (no. of tyrosine hydroxylase positive cells (TH+ cells) in the substantia nigra pars compacta (SNpc)) and functionality (dopamine and its metabolites DOPAC and HVA content in the striata) of nigrostriatal dopaminergic neurons were assessed. In the vehicle‐treated rats, infusion of 8 μg of 6‐OHDA significantly reduced the number of TH+ cells in the SNpc as well as the levels of dopamine, DOPAC and HVA in the striata on the lesion side. The loss of TH+ cells, dopamine and HVA, but not the DOPAC levels, was significantly attenuated by stdEM‐X pretreatment, but not by the dilEM‐X pretreatment. There were no significant changes in the TH+ cells, or in the monoamine levels with the EM‐X pretreatment per se, except for a small but significant fall in the levels of dopamine with the stdEM‐X. The evidence presented supports the potential neuroprotective effects of stdEM‐X drink, although its effect on dopamine levels needs further investigation.

[1]  Minoru Harada,et al.  Tumor necrosis factor-α (TNF-α) increases both in the brain and in the cerebrospinal fluid from parkinsonian patients , 1994, Neuroscience Letters.

[2]  Fumitaka Osakada,et al.  Neuroprotective effects of α-tocopherol on oxidative stress in rat striatal cultures , 2003 .

[3]  C. Marsden,et al.  Oxidative stress as a cause of nigral cell death in Parkinson's disease and incidental lewy body disease , 1992, Annals of neurology.

[4]  H. Chapel,et al.  CLINICAL RELEVANCE OF SPECIFIC IgG ANTIBODIES TO CARDIOLIPIN , 1989, The Lancet.

[5]  P. Pietta,et al.  Flavonoids as antioxidants. , 2000, Journal of natural products.

[6]  D. Dexter,et al.  Chronic L‐DOPA administration is not toxic to the remaining dopaminergic nigrostriatal neurons, but instead may promote their functional recovery, in rats with partial 6‐OHDA or FeCl3 nigrostriatal lesions , 2001, Movement disorders : official journal of the Movement Disorder Society.

[7]  S. Mandel,et al.  Attenuation of 6-hydroxydopamine (6-OHDA)-induced nuclear factor-kappaB (NF-kappaB) activation and cell death by tea extracts in neuronal cultures. , 2002, Biochemical pharmacology.

[8]  R. Nicolosi,et al.  Efficacy of vitamin E, phosphatidyl choline, and pyruvate on buffering neuronal degeneration and oxidative stress in cultured cortical neurons and in central nervous tissue of apolipoprotein E-deficient mice. , 2002, Free radical biology & medicine.

[9]  B. Steinmann,et al.  Mass fragmentography of dopamine and 6-hydroxydopamine. Application to the determination of dopamine in human brain biopsies from the caudate nucleus. , 1974, Journal of chromatography.

[10]  N. Holbrook,et al.  Oxidants, oxidative stress and the biology of ageing , 2000, Nature.

[11]  A. Izzo,et al.  Flavonoids: old and new aspects of a class of natural therapeutic drugs. , 1999, Life sciences.

[12]  A. Bast,et al.  Influence of iron chelation on the antioxidant activity of flavonoids. , 1998, Biochemical pharmacology.

[13]  L Bravo,et al.  Polyphenols: chemistry, dietary sources, metabolism, and nutritional significance. , 2009, Nutrition reviews.

[14]  R. Burke,et al.  6-Hydroxydopamine lesion of the rat substantia nigra: time course and morphology of cell death. , 1995, Neurodegeneration : a journal for neurodegenerative disorders, neuroprotection, and neuroregeneration.

[15]  O. Aruoma,et al.  Assessment of the ability of the antioxidant cocktail-derived from fermentation of plants with effective microorganisms (EM-X) to modulate oxidative damage in the kidney and liver of rats in vivo: studies upon the profile of poly- and mono-unsaturated fatty acids. , 2002, Toxicology letters.

[16]  P. Riederer,et al.  Tumor necrosis factor-alpha (TNF-alpha) increases both in the brain and in the cerebrospinal fluid from parkinsonian patients. , 1994, Neuroscience letters.

[17]  Ke Bin.  Antiaging Activities of Effective Microorganisms Extract(EM- X) on the Fruit Fly and the Mouse , 2001 .

[18]  A. Gomes,et al.  Antiinflammatory and antioxidant property of saponins of tea [Camellia sinensis (L) O. Kuntze] root extract , 2001, Phytotherapy research : PTR.

[19]  D. Calne,et al.  AETIOLOGY OF PARKINSON'S DISEASE , 1983, The Lancet.

[20]  A. Schapira,et al.  Mitochondrial involvement in Parkinson’s disease , 2002, Neurochemistry International.

[21]  O. Aruoma,et al.  Antioxidant activities of phenolic, proanthocyanidin, and flavonoid components in extracts of Cassia fistula. , 2002, Journal of agricultural and food chemistry.

[22]  Stanley J. Watson,et al.  The rat brain in stereotaxic coordinates (2nd edn) by George Paxinos and Charles Watson, Academic Press, 1986. £40.00/$80.00 (264 pages) ISBN 012 547 6213 , 1987, Trends in Neurosciences.

[23]  A. Collins,et al.  Quercetin and myricetin protect against hydrogen peroxide-induced DNA damage (strand breaks and oxidised pyrimidines) in human lymphocytes. , 1997, Mutation research.

[24]  M. Tirmenstein,et al.  Vitamin E succinate protects hepatocytes against the toxic effect of reactive oxygen species generated at mitochondrial complexes I and III by alkylating agents. , 2001, Chemico-biological interactions.

[25]  I. Shoulson.  DATATOP: a decade of neuroprotective inquiry. Parkinson Study Group. Deprenyl And Tocopherol Antioxidative Therapy Of Parkinsonism. , 1998, Annals of neurology.

[26]  R. Andrew,et al.  The determination of hydroxydopamines and other trace amines in the urine of Parkinsonian patients and normal controls , 1993, Neurochemical Research.

[27]  C. Marsden,et al.  Basal Lipid Peroxidation in Substantia Nigra Is Increased in Parkinson's Disease , 1989, Journal of neurochemistry.

[28]  C. Marsden,et al.  Alterations in glutathione levels in Parkinson's disease and other neurodegenerative disorders affecting basal ganglia , 1994, Annals of neurology.

[29]  S A van Acker,et al.  Structural aspects of antioxidant activity of flavonoids. , 1996, Free radical biology & medicine.

[30]  N. Sugihara,et al.  Anti- and pro-oxidative effects of flavonoids on metal-induced lipid hydroperoxide-dependent lipid peroxidation in cultured hepatocytes loaded with alpha-linolenic acid. , 1999, Free radical biology & medicine.

[31]  F. L. Muiswinkel,et al.  Drug treatment of Parkinson's disease. Time for phase II. , 2000, Biochemical pharmacology.

[32]  A. Korczyn.  Drug treatment of Parkinson's disease , 2004, Dialogues in clinical neuroscience.

[33]  N. Quinn,et al.  Fortnightly Review: Drug treatment of Parkinson's disease , 1995 .

[34]  O. Aruoma.  Characterization of drugs as antioxidant prophylactics. , 1996, Free radical biology & medicine.

[35]  Ira Shoulson,et al.  DATATOP: A decade of neuroprotective inquiry , 1998 .

[36]  C. Marsden,et al.  INCREASED NIGRAL IRON CONTENT IN POSTMORTEM PARKINSONIAN BRAIN , 1987, The Lancet.

[37]  J. Joseph,et al.  Structure-activity relationships of quercetin in antagonizing hydrogen peroxide-induced calcium dysregulation in PC12 cells. , 1999, Free radical biology & medicine.

[38]  T. Higa,et al.  The antioxidant cocktail effective microorganism X (EM-X) inhibits oxidant-induced interleukin-8 release and the peroxidation of phospholipids in vitro. , 2002, Biochemical and biophysical research communications.

[39]  D. Dexter,et al.  Tissue distribution and neuroprotective effects of citrus flavonoid tangeretin in a rat model of Parkinson's disease , 2001, Neuroreport.

[40]  S. Fahn.  A pilot trial of high‐dose alpha‐tocopherol and ascorbate in early Parkinson's disease , 1992, Annals of neurology.

[41]  G. Paxinos,et al.  The Rat Brain in Stereotaxic Coordinates , 1983 .